Stevia Composition

ABSTRACT

Stevia  compositions are prepared from steviol glycosides of  Stevia rebaudiana  Bertoni. The compositions are able to provide a superior taste profile and can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals.

BACKGROUND OF THE INVENTION Field of the Invention

The invention relates to a process for producing a highly purified foodingredient from the extract of the Stevia rebaudiana Bertoni plant andits use in various food and beverage products.

Description of the Related Art

Nowadays sugar alternatives are receiving increasing attention due toawareness of many diseases in conjunction with consumption of high-sugarfoods and beverages. However many artificial sweeteners such as dulcin,sodium cyclamate and saccharin were banned or restricted in somecountries due to concerns on their safety. Therefore non-caloricsweeteners of natural origin are becoming increasingly popular. Thesweet herb Stevia rebaudiana Bertoni produces a number of diterpeneglycosides which feature high intensity sweetness and sensory propertiessuperior to those of many other high potency sweeteners.

The above-mentioned sweet glycosides, have a common aglycon, steviol,and differ by the number and type of carbohydrate residues at the C13and C19 positions. The leaves of Stevia are able to accumulate up to10-20% (on dry weight basis) steviol glycosides. The major glycosidesfound in Stevia leaves are Rebaudioside A (2-10%), stevioside (2-10%),and Rebaudioside C (1-2%). Other glycosides such as Rebaudioside B, D,E, and F, steviolbioside and rubusoside are found at much lower levels(approx. 0-0.2%).

Two major glycosides—stevioside and Rebaudioside A (Reb A), wereextensively studied and characterized in terms of their suitability ascommercial high intensity sweeteners. Stability studies in carbonatedbeverages confirmed their heat and pH stability (Chang S. S., Cook, J.M. (1983) Stability studies of stevioside and Rebaudioside A incarbonated beverages. J. Agric. Food Chem. 31: 409-412.)

Steviol glycosides differ from each other not only by molecularstructure, but also by their taste properties. Usually stevioside isfound to be 110-270 times sweeter than sucrose, Rebaudioside A between150 and 320 times, and Rebaudioside C between 40-60 times sweeter thansucrose. Dulcoside A is 30 times sweeter than sucrose. Rebaudioside Ahas the least astringent, the least bitter, and the least persistentaftertaste thus possessing the most favorable sensory attributes inmajor steviol glycosides (Tanaka O. (1987) Improvement of taste ofnatural sweeteners. Pure Appl. Chem. 69:675-683; Phillips K. C. (1989)Stevia: steps in developing a new sweetener. In: Grenby T. H. ed.Developments in sweeteners, vol. 3. Elsevier Applied Science, London.1-43). The chemical structure of Rebaudioside A is shown in FIG. 1.

Methods for the extraction and purification of sweet glycosides from theStevia Rebaudiana plant using water or organic solvents are describedin, for example, U.S. Pat. Nos. 4,361,697; 4,082,858; 4,892,938;5,972,120; 5,962,678; 7,838,044 and 7,862,845.

However, even in a highly purified state, steviol glycosides stillpossess non-sweet taste attributes such as bitterness, sweet aftertaste,licorice flavor, etc. One of the main obstacles for the successfulcommercialization of Stevia sweeteners are such non-sweet tasteattributes. It was shown that these flavor notes become more prominentas the concentration of steviol glycosides increases (Prakash I., DuBoisG. E., Clos J. F., Wilkens K. L., Fosdick L. E. (2008) Development ofRebiana, a natural, non-caloric sweetener. Food Chem. Toxicol., 46,S75-S82).

Rebaudioside B (CAS No: 58543-17-2), or Reb B, also known as steviosideA₄ (Kennelly E. J. (2002) Constituents of Stevia rebaudiana In Stevia:The genus Stevia, Kinghorn A. D. (Ed), Taylor & Francis, London, p. 71),is one of the sweet glycosides found in Stevia rebaudiana. Sensoryevaluations show that Reb B was approximately 300-350 times sweeter thansucrose, while for Reb A this value was approximately 350-450 (Crammer,B. and Ikan, R. (1986) Sweet glycosides from the Stevia plant. Chemistryin Britain 22, 915-916, and 918). The chemical structure of RebaudiosideB is shown in FIG. 2 a.

It was believed that Reb B forms from partial hydrolysis of RebaudiosideA during the extraction process (Kobayashi, M., Horikawa, S., Degrandi,I. H., Ueno, J. and Mitsuhashi, H. (1977) Dulcosides A and B, newditerpene glycosides from Stevia rebaudiana. Phytochemistry 16,1405-1408). However further research shows that Reb B occurs naturallyin the leaves of Stevia rebaudiana and currently it is one of ninesteviol glycosides recognized by FAO/JECFA (United Nations' Food andAgriculture Organization/Joint Expert Committee on Food Additives) incalculating total steviol glycosides' content in commercial steviolglycosides preparations (FAO JECFA (2010) Steviol Glycosides, Compendiumof Food Additive Specifications, FAO JECFA Monographs 10, 17-21).

It is also noted that no significant work has been conducted todetermine the potential of Reb B as a sweetener or food ingredient.Moreover Reb B is often viewed as process artifact and unnecessaryimpurity in commercial steviol glycosides preparations. No significantevaluation of Reb B influence on overall taste profile of steviolglycosides preparations has been conducted.

In food and beverage applications, the solubility of high intensitysweeteners like steviol glycosides is very important and can be asignificant barrier to achieve the desirable sweetness and tasteprofile. However highly purified steviol glycosides possess relativelylow water solubility. For example Rebaudioside A (Reb A) thermodynamicequilibrium solubility at room temperature is only 0.8%. On the otherhand, the water solubility of Reb B is reported to be about 0.1% andthat of Reb D (FIG. 2b ) is even lower, only at 0.01-0.05% at roomtemperature (Kinghorn A. D. (2002) Constituents of Stevia rebaudiana InStevia: The genus Stevia, Kinghorn A. D. (Ed), Taylor & Francis, London,p. 8). Reb B or Reb D can be solubilized at higher concentrations at ahigher temperature, but they will quickly crystallize back from solutionupon cooling to room temperature. Considering high sweetness intensityof steviol glycosides (100-300 times of sugar sweetness)—even 0.05%solubility may seem sufficient for many applications.

In many food processes where highly concentrated ingredients are used, ahighly soluble form of Reb B and Reb D will be necessary.

Considering the facts mentioned above, it is necessary to evaluate Reb Bas a sweetener and food ingredient and develop a simple and efficientprocess for food grade Reb B preparations suitable for foodapplications.

SUMMARY OF THE INVENTION

In this invention, a process is developed to prepare combinations ofdifferent glycosides and food ingredients with Reb B. The process allowsto preparation of blends more soluble than a mechanical blend of initialingredients.

Within the description of this invention we show that, when applied inspecific manner, certain blend of Reb B with other steviol glycosides,may impact the taste profile and offer significant advantages for Steviasweeteners' use in various applications.

In an embodiment of the present invention, certain blend of Reb B withReb A may impact the taste profile and offer significant advantages forStevia sweeteners' use in various applications.

In one embodiment of present invention certain blend of Reb B with RebD, impacts the taste profile and offer significant advantages for Steviasweeteners' use in various applications.

In another embodiment of present invention we show that, when applied inspecific manner, certain blend of Reb B and other steviol glycosideswith sweeteners from sugar alcohol group impact the taste profile andoffer significant advantages for Stevia sweeteners' use in variousapplications.

In yet another embodiment of present invention we show that, whenapplied in specific manner, certain blend of Reb B and other steviolglycosides with sweeteners selected from group of natural high intensitysweetener impact the taste profile and offer significant advantages forStevia sweeteners' use in various applications.

In one embodiment of present invention we show that, when applied inspecific manner, certain blend of Reb B and other steviol glycosideswith compositions selected from glycosylated natural high intensitysweetener group impact the taste profile and offer significantadvantages for Stevia sweeteners' use in various applications.

In another embodiment of present invention we show that, when applied inspecific manner, certain blend of Reb B and other steviol glycosideswith sweeteners selected from group of synthetic high intensitysweeteners impact the taste profile and offer significant advantages forStevia sweeteners' use in various applications.

In yet another embodiment of present invention we show that, whenapplied in specific manner, certain blend of Reb B and other steviolglycosides with ingredients selected from oligosaccharide group impactthe taste profile and offer significant advantages for Steviasweeteners' use in various applications.

In one embodiment of present invention we show that, when applied inspecific manner, certain blend of Reb B and other steviol glycosideswith sweeteners from caloric sweetener group impact the taste profileand offer significant advantages for Stevia sweeteners' use in variousapplications.

Hereinafter the term “steviol glycoside(s)” will mean Rebaudioside A(Reb A), Rebaudioside B (Reb B), Rebaudioside C (Reb C), Rebaudioside D(Reb D), Rebaudioside E (Reb E), Rebaudioside F (Reb F), Stevioside(Stev), Steviolbioside (Sbio), Dulcoside A (Dulc A), Rubusoside (Rub),or other glycoside of steviol and/or combinations thereof.

Hereinafter the terms “total steviol glycosides”, or “total glycosides”,or “TSG”, will mean the sum of concentrations (% wt/wt on anhydrousbasis) of Reb A, Reb B, Reb C, Reb D, Reb E, Reb F, Stevioside,Steviolbioside, Dulcoside A, Rubusoside, or other glycosides of steviol.

Hereinafter unless specified otherwise the purity of used Reb A, Reb B,and Reb D, is at least 95% (wt/wt, on anhydrous basis).

Hereinafter the terms “Reb A/B” and “Reb A/Reb B” will meanblends/mixtures of Reb A and Reb B prepared by process of presentinvention.

Hereinafter, unless specified otherwise the solubility of material isdetermined in reverse osmosis (RO) water at room temperature. Where thesolubility is expressed as “%” it to be understood as number of grams ofmaterial soluble in 100 grams of solvent.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory and areintended to provide further explanation of the invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are included to provide a furtherunderstanding of the invention. The drawings illustrate embodiments ofthe invention and together with the description serve to explain theprinciples of the embodiments of the invention.

FIG. 1 shows the chemical structure of Reb A.

FIGS. 2a and 2b show the chemical structures of Reb B and Reb D,respectively.

FIG. 3 shows the sweetness profile of compositions featuring differentratios of Reb A to Reb B (Reb A/Reb B).

FIG. 4 shows the effect of Reb B and a blend of steviol glycosides onsweetness profile of Mogroside in water.

FIGS. 5a and 5b show the effect of Reb B and a blend of steviolglycosides on sweetness profile of Sucralose in water and in a flavoreddrink.

FIGS. 6a and 6b show the effect of Reb B and a blend of steviolglycosides on sweetness profile of Erythritol in water and in a flavoreddrink.

FIGS. 7a and 7b show the effect of Reb B and a blend of steviolglycosides on sweetness profile of FOS or Fructooligosaccharide in waterand in an alcoholic beverage.

DETAILED DESCRIPTION OF THE INVENTION

Advantages of the present invention will become more apparent from thedetailed description given hereinafter. However, it should be understoodthat the detailed description and specific examples, while indicatingpreferred embodiments of the invention, are given by way of illustrationonly, since various changes and modifications within the spirit andscope of the invention will become apparent to those skilled in the artfrom this detailed description.

In one embodiment of invention a process is developed to preparecombinations of different glycosides and food ingredients with Reb B.

In one embodiment the invention describes a sweetened ingestible productcomprising blends of Reb B and at least one sweetener, selected fromdifferent groups of sweet molecules for producing a blend of sweetenersand use thereof in various food and beverage products as a sweetener andflavor modifier. The groups of sweet molecules comprise and includecombinations of the following:

-   -   Steviol glycosides and Stevia extracts,    -   Other natural sweeteners (Luo Han Guo extract, Mogrosides,        Mogroside V, Glycyrrhizin, Thaumatin, Brazzein, etc),    -   Glycosylated terpenoid sweeteners (glucosylated steviol        glycosides and glucosylated mogrosides, etc)    -   Synthetic high intensity sweeteners (aspartame, saccharin,        sucralose, Acesulfame-K, etc),    -   Sugar alcohols (erythritol, maltitol, sorbitol, etc),    -   Oligosaccharides (fructooligosaccharides, inulin,        inulooligosaccharides, maltooligosaccharides etc) and    -   Caloric sweeteners (sugar, fructose, dextrose, maltose, lactose,        corn syrup, and HFCS etc).

The invention, in part, pertains to an ingredient comprising selectedmolecules of steviol glycosides (as defined above) of Stevia rebaudianaBertoni plant.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and other steviol glycosides.

The invention, in part, pertains to an ingredient comprising steviolglycosides of Stevia rebaudiana Bertoni plant and a sweetener (orcombination of sweeteners) from other natural sweeteners including butnot limited to, Luo Han Guo extract, Mogrosides, Mogroside V,Glycyrrhizin, Thaumatin, Brazzein, etc.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and a sweetener (or combinationof sweeteners) from other natural sweeteners including but not limitedto, Luo Han Guo extract, Mogrosides, Mogroside V, Glycyrrhizin,Thaumatin, Brazzein, etc.

The invention, in part, pertains to ingredients comprising steviolglycosides of Stevia Rebaudiana Bertoni plant and a compositioncomprising glycosylated terpenoid sweeteners including but not limitedto, glucosylated Steviol Glycosides and glucosylated Mogrosides.

In one embodiment the invention, in part, pertains to ingredientscomprising specific blends of steviol glycosides and Reb B, and acomposition comprising glycosylated terpenoid sweeteners including butnot limited to, Glucosylated Steviol Glycosides and GlucosylatedMogrosides.

The invention, in part, pertains to ingredients comprising steviolglycosides of Stevia Rebaudiana Bertoni plant and a sweetener (orcombination of sweeteners) from synthetic high intensity sweetenersincluding but not limited to aspartame, saccharin, sucralose,Acesulfame-K, neotame.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and a sweetener (or combinationof sweeteners) from synthetic high intensity sweeteners including butnot limited to aspartame, saccharin, sucralose, Acesulfame-K, neotame.

The invention, in part, pertains to an ingredient comprising steviolglycosides of Stevia rebaudiana Bertoni plant and a sweetener (orcombination of sweeteners) from sugar alcohols including but not limitedto erythritol, maltitol, sorbitol, xylitol.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and a sweetener (or combinationof sweeteners) from sugar alcohols including but not limited toerythritol, maltitol, sorbitol, xylitol.

The invention, in part, pertains to an ingredient comprising steviolglycosides of Stevia rebaudiana Bertoni plant and a sweetener (orcombination of sweeteners) from oligosaccharides including but notlimited to fructo-oligosaccharides, inulin, inulo-oligosaccharides,polydextrose, and malto-oligosaccharides.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and a sweetener (or combinationof sweeteners) from oligosaccharides including but not limited tofructo-oligosaccharides, inulin, inulo-oligosaccharides, polydextrose,and malto-oligosaccharides.

The invention, in part, pertains to an ingredient comprising steviolglycosides of Stevia rebaudiana Bertoni plant and a sweetener (orcombination of sweeteners) from caloric sweeteners group including butnot limited to sugar, invert-sugar, fructose, dextrose, maltose,lactose, corn syrup, and HFCS.

The invention, in part, pertains to ingredients comprising specificblends of steviol glycosides and Reb B, and a sweetener (or combinationof sweeteners) from caloric sweeteners group including but not limitedto sugar, invert-sugar, fructose, dextrose, maltose, lactose, cornsyrup, and HFCS.

In another aspect, a sweetened ingestible product comprising a) at leastone sweetener, selected from the group consisting of: steviolglycosides, other natural sweeteners, glycosylated terpenoid sweeteners,synthetic high intensity sweeteners, sugar alcohols, oligosaccharides,caloric sweeteners, and a combination thereof, wherein said at least onesweetener or sweetener combination is present in the product at aconcentration above 2% sucrose equivalent sweetness, and b) Reb B at aconcentration from 10 to 300 ppm.

In another aspect, the invention is also directed to a method forproducing a soluble sweetener composition, comprising the steps ofproviding low solubility Stevia sweeteners, solubilizing them in waterunder gradient temperature treatment conditions, to produce highlystable concentrated solution, and spray drying the highly stableconcentrated solution to obtain a highly soluble Stevia composition.

In one embodiment of this invention the initial materials for preparingsoluble sweetener composition were selected from the group including RebD, Reb A, Reb B, and steviolbioside (Sbio).

In another aspect of this invention, steviol glycosides compositionswith lower solubility, (lower than 1%) were combined with Reb B throughthe method described above, and yielded compositions with significantlyhigher solubility (more than 1%). This phenomenon was unexpected, aspure Reb B on its own has <0.1% solubility.

The compositions of present invention can be used as sweetness enhancer,flavor enhancer and sweetener in various food and beverage products.Non-limiting examples of food and beverage products include carbonatedsoft drinks, ready to drink beverages, energy drinks, isotonic drinks,low-calorie drinks, zero-calorie drinks, sports drinks, teas, fruit andvegetable juices, juice drinks, flavored water, dairy drinks, yoghurtdrinks, alcohol beverages, powdered beverages, bakery products, bread,cookies, biscuits, muffins, rolls, baking mixes, cereals, breakfastcereals confectioneries, candies, toffees, chewing gum, frostings, dairyproducts, flavored milk, yoghurts, flavored yoghurts, cultured milk,frozen dairy desserts including ice cream, sauces and gravies, soy sauceand other soy base products, salad dressings, mayonnaise, vinegar,condiments and relishes, meat products, fish-meat products, bottled andcanned foods, frozen-desserts, jams and jellies, gelatins, puddings andfillings tabletop sweeteners, processed fruits and vegetables.

Additionally the compositions can be used in drug or pharmaceuticalpreparations and cosmetics, including but not limited to toothpaste,mouthwash, cough syrup, chewable tablets, lozenges, vitaminpreparations, and the like.

The compositions can be used “as-is” or in combination with othersweeteners, flavors and food ingredients.

Non-limiting examples of sweeteners include steviol glycosides,stevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, RebaudiosideD, Rebaudioside E, Rebaudioside F, dulcoside A, steviolbioside,rubusoside, as well as other steviol glycosides found in Steviarebaudiana Bertoni plant and mixtures thereof, Stevia extract, Luo HanGuo extract, mogrosides, high-fructose corn syrup, corn syrup, invertsugar, fructooligosaccharides, inulin, inulooligosaccharides, couplingsugar, maltooligosaccharides, maltodextins, corn syrup solids, glucose,maltose, sucrose, lactose, aspartame, saccharin, sucralose, sugaralcohols.

Non-limiting examples of flavors include lemon, orange, fruit, banana,grape, pear, pineapple, bitter almond, cola, cinnamon, sugar, cottoncandy, vanilla flavors.

Non-limiting examples of other food ingredients include flavors,acidulants, organic and amino acids, coloring agents, bulking agents,modified starches, gums, texturizers, preservatives, antioxidants,emulsifiers, stabilizers, thickeners, gelling agents.

The following examples illustrate various embodiments of the invention.It will be understood that the invention is not limited to thematerials, proportions, conditions and procedures set forth in theexamples, which are only illustrative.

Example 1 Preparation of Reb D and Reb B Soluble Composition

70 g of rebaudioside D with 98.1% purity (dry weight basis), havingwater solubility of 0.03% and 30 g of rebaudioside B with 99.0% purity(dry weight basis), and having water solubility of 0.01%, both producedby PureCircle Sdn Bhd (Malaysia), were mixed with 400 g water andincubated in thermostatted oil bath. The temperature was increased at 1°C. per minute to 121° C. The mixture was maintained at 121° C. for 1hour and then the temperature was decreased to 80° C., at 1° C. perminute, and the obtained solution was spray dried using YC-015laboratory spray drier (Shanghai Pilotech Instrument & Equipment Co.Ltd., China) operating at 175° C. inlet and 100° C. outlet temperature.Solution was maintained at 80° C. to prevent premature crystallization.About 90 g of amorphous powder was obtained with 1% solubility.

Example 2 Effect of Stevia Composition on Solubility in Water

Solubility were evaluated for Reb A, Reb B, mechanical blends of Reb Aand Reb B powders of different ratios, and mixtures of Reb A and Reb Bprepared according to process described in EXAMPLE 1. Reb A and Reb Bshowed solubility around 1% and 0.1% at room temperature and increasedsolubility on heating to a higher temperature. Table 1 shows thesolubility of different ingredients and their blends.

TABLE 1 Reb A/Reb B blends (wt/wt ratio of Reb B to total glycosides)Blend made by Solubility Pure Pure Mechanical blend process of inventionin water Reb A Reb B 10% 16% 26% 10% 16% 26% 0.5%   Yes No* No* No* No*Yes Yes Yes 1% Yes No* No* No* No* Yes Yes Yes 2% No No* No* No* No* YesYes Yes 5% No No* No* No* No* Yes Yes Yes *Suspension was opaque andparticles settled down when mixing was stopped

Sweetness Factor (SF) Determination

The sweetness factors for different sweetener from all major groups weremeasured by tasting several concentrations of each sweetener in water bytrained panel. From the sensory test at a, the concentration (%) thatcorresponds to the 5% sucrose equivalent (SE) sweetness were estimatedand listed in Table 2. The sweetness equivalent (SE) of high intensitysweeteners (HIS) varies according to the target sugar equivalentsweetness levels, that are tested as shown in Table 3. Table 3 alsoshows the sweetness factors of Reb A, Reb B and a test sample (Reb A/B)with a blend of Reb A and Reb Bat 84:16 ratio.

TABLE 2 Sweetness factors of selected sweeteners Sweetener UsageCalculated Group Sweetener Level, % SF Steviol Glycoside Reb A (97%)0.0172 291 Reb D (97%) 0.0172 291 Reb B (98%) 0.0397 126 Other NaturalHIS Mogroside V (70%) 0.0212 236 Glycosylated Glucosylated 0.0556 90Terpenoid sweetener Steviol Glycosides Synthetic HIS Sucralose 0.0083602 Sugar Alcohol Erythritol 7.49 0.67 Maltitol 6.44 0.77Oligosaccharide FOS (Frutalose L90) 21 0.24 Caloric Sweetener Sugar 5 1

TABLE 3 Sweetness factor of selected steviol glycosides SE levels 2.5%5.0% 7.5% 10.0% Reb A 400 290 250 200 Reb B 146 126 95 59 Reb A/B 350260 220 190

Example 4

Effect of Stevia Composition on Sweetness Profile in Water

In another embodiment, Stevia sweetener (Reb A, Reb B or blends thereof)solutions were made by completely dissolving Stevia into distilledwater. Solutions were tasted and evaluated by a trained panel. FIG. 3shows the effect of different ratio of Reb A and Reb Bon sweetnessprofile and associated aftertastes (vertical axis is a relativeintensity of different attributes). All solutions were made withdifferent mass ratio of Reb A and Reb Bas shown in FIG. 3 to get the 7%sweetness equivalent. Reb B a mount varied between 20 mg/kg to 150mg/kg. It is apparent that the blend of Reb A and Reb B showunexpectedly improved sweetness profile at certain ratios.

Example 5

In one embodiment, Reb B was combined (at 50, 150 and 300 ppm) withseveral sweeteners listed in Table 2 to make solutions in plain water.Some of the sweeteners were tested at two different concentrations toobtain around 2% sugar equivalent sweetness without any Reb B. A trainedpanel conducted sensory tests to determine the effect of the addition ofdifferent amounts of Reb B to each sweetener on overall sweetnessquality and intensity as shown in Table 4. In presence of Reb B, most ofthe sweeteners showed a surprising level of enhancement of sweetnessintensity. The addition of Reb B generally increased upfront sweetness,made the sweetness profile rounded and contributed to sweetnesslingering at a higher level.

TABLE 4 Sugar equivalent sweetness (%) for different sweetenercombinations SE of Sweetener/Reb B combination 0 ppm 50 ppm 150 ppm 300ppm Sweetener Reb B Reb B Reb B Reb B Mogroside V (100 ppm)   2.5 4  5.5 8-8.5 Mogroside V (212 ppm) 5.5-6    7 9 10-10.5 Sucralose (40ppm) 2-2.5 5 7.5-8 11 Sucralose 83 ppm 5 7 11  14 Erythritol (3.5%)1-1.5 3 5.5-6 9 Erythritol (5.0%) 3 4 8 9.5 Maltitol (6.44%) 5 6 9 12FOS (21%) 5 6.5 8 11

Example 6

In one embodiment, several concentrations of the Reb A/B blend (used inEXAMPLE 3) were tested to match the sweetness of Reb B solutions at 50,150 and 300 ppm. A trained panel of 13 members found that 22, 68 and 128ppm of Reb A/B matched the sweetness intensity of Reb B solutions with50, 150 and 300 ppm concentrations respectively. Table 5 shows thecombination of different sweeteners that were used for differentapplications to show the effect of Reb B and Reb A/B on the sweetnessand flavor profile in different applications.

All sensory attributes were rated by trained panelists on a scale ofzero (0) to ten (10). Samples were presented in a randomized fashion andlabeled with three-digit codes.

Sweet Onset—This attribute describes the time at which panelistsperceived sweetness upon placing the sample in the mouth. A score ofzero indicates immediate identification of sweetness, whereas a score often indicates a long delay in the time that sweetness was perceived.

Total Sweetness—This attribute describes the highest intensity ofsweetness perceived during tasting. A score of zero indicates nosweetness, while a score of ten indicates very high sweetness.

Bitterness—This attribute describes the highest intensity of bitternessperceived during tasting. A score of zero indicates no bitterness, whilea score of ten indicates very high bitterness.

Rounded Sweetness—This attribute describes the duration of sweetnesswhile samples are in the mouth. A score of zero indicates that thesweetness comes and goes very quickly and is more like high intensitysweeteners in its temporal profile, while a score of ten indicates thatthe sweetness lasts for a long time, and is more similar to sugar in itstemporal profile.

Flavor Intensity—This attribute describes the highest intensity offlavor perceived during tasting. A score of zero indicates no flavor,while a score of ten indicates very high flavor.

Astringency—This attribute describes the highest intensity ofastringency perceived during tasting. A score of zero indicates noastringency, while a score of ten indicates very high astringency.

Mouth Coating—This attribute describes the highest intensity ofsweetness coating perceived while samples are in the mouth. A score ofzero indicates that there is no coating of sweetness, while a score often indicates very high sweetness coating.

Sweet Lingering—This attribute describes the highest intensity ofsweetness perceived after the sample has been swallowed. A score of zeroindicates that there is no sweetness perceived after the sample has beenswallowed, while a score of ten indicates high sweetness after thesample has been swallowed.

Bitter Lingering—This attribute describes the highest intensity ofbitterness perceived after the sample has been swallowed. A score ofzero indicates that there is no bitterness perceived after the samplehas been swallowed, while a score of ten indicates high bitterness afterthe sample has been swallowed.

Aftertaste Mouth Coating—This attribute describes the highest intensityof sweetness coating perceived after the sample has been swallowed. Ascore of zero indicates that there is no sweetness coating perceivedafter the sample has been swallowed, while a score of ten indicates highsweetness coating after the sample has been swallowed.

Tartness—This attribute describes the highest intensity of tartnessperceived during tasting. A score of zero indicates no tartness, while ascore of ten indicates very high tartness.

Cooling Sensation—This attribute describes the highest intensity ofcooling sensation perceived during tasting. A score of zero indicates nocooling sensation, while a score of ten indicates very high coolingsensation.

The consideration of these attributes was used for selecting differentsweetener combinations for application examples.

TABLE 5 Sweetener combination tested for applications SweetenerSweetener + Reb B Sweetener + Reb A/B Erythritol 5% +50 ppm +22 ppmSucralose 40 ppm +150 ppm +68 ppm Mogroside V 100 pm +300 ppm +128 ppmMaltitol (6.44%) +50 ppm +22 ppm FOS (21%) +150 ppm +68 ppm

Example 7

Water solution of mogroside (100 ppm) sweetener was tested againstsolutions made with mogroside (100 ppm)+Reb B (300 ppm) and mogroside(100 ppm)+Reb A/B (Example 3) blend (128 ppm) by a 13 members trainedpanel. They found Reb B with mogroside sample increases sweetness levelsignificantly, improves the onset of sweetness and decreases thebitterness. In presence of Reb A/B blend and mogroside, the increase insweetness is higher than mogroside solution, but lower thanmogroside+Reb B solution. FIG. 4 shows the difference in tasteattributes for all three sweetener systems.

Example 8

Water solution of sucralose (40 ppm) was tested against solutions madewith sucralose (40 ppm)+Reb B (150 ppm) and sucralose (40 ppm)+Reb A/RebB (Example 3) blend (68 ppm) by a 13 members trained panel. They foundReb B with sucralose sample had significantly higher sweetness level.Reb B improved the onset of sweetness and decreased the off tasteslightly. In presence of Reb A/Reb B blend and sucralose, the increasein sweetness is higher than sucralose solution, but lower thansucralose+Reb B solution. FIG. 5a shows the difference in tasteattributes for all three sweetener systems containing sucralose andsteviol glycosides.

Example 9

The sucralose sweetener and the steviol glycosides compositions(described in EXAMPLE 8) were used for making a mango-passion fruitflavored Powder Soft Drink (PSD) and a sensory panel found the softdrink containing sucralose with steviol glycosides significantlyincreased the sweetness, enhanced the fruit flavor, reduced theastringency, and made the drink more rounded in sweetness and acidprofile. FIG. 5b shows the impact of different sweetener system on thetaste attributes of the beverage.

Example 10

Water solution of erythritol (5%) was tested against solutions made witherythritol (5%) +Reb B (50 ppm) and erythritol (5%) +Reb A/Reb B(Example 3) blend (22 ppm) by a 13 members trained panel. They found RebB improved the taste of erythritol sample significantly by increasingthe sweetness and reducing the bitterness and cooling effect. Along withhigher sweetness level, Reb B improved the onset of sweetness and thepeak sweetness without increasing the sweetness lingering (sweet aftertaste). In presence of Reb A/Reb B blend and erythritol, the solutionhad a higher sweetness and showed fairly similar improvement of otherattributes as Reb B showed with erythritol. FIG. 6a shows the differencein taste attributes for all three sweetener systems containing sucraloseand steviol glycosides.

Example 11

The erythritol sweetener and the steviol glycosides compositions(described in EXAMPLE 10) were used for making a mango-passion fruitflavored Powder Soft Drink (PSD) and a sensory panel found the softdrink containing erythritol with steviol glycosides significantlyincreased the sweetness, reduced the bitterness, enhanced the fruitflavor, reduced the astringency, and made the drink more rounded insweetness and acid profile. Steviol glycosides also reduced theaftertaste. FIG. 6b shows the impact of different sweetener systemcontaining erythritol on the taste attributes of the beverage.

Example 12

Fructo-oligosaccharide or FOS solution (21%) was tested againstsolutions made with FOS (21%)+Reb B (150 ppm) and FOS (21%)+Reb A/Reb B(Example 3) blend (68 ppm) by a 13 members trained panel. They found RebB with FOS sample had significantly higher sweetness and made thesweetness profile rounded. In presence of Reb A/Reb B blend and FOS, theincrease in sweetness is higher than FOS solution but lower thansucralose+Reb B solution. FIG. 7a shows the difference in tasteattributes for all three sweetener systems containing FOS and steviolglycosides.

Example 13

The Fructo-oligosaccharide (FOS) sweetener and the steviol glycosidescompositions (described in EXAMPLE 12) were used for making an alcoholicbeverage—lime-tequila margarita drink using Margarita pre-mix from FIS(Flavor International Systems), recommended usage was 0.4%. PreparedLime Tequila Margarita blank containing about 10% alcohol, no sugaradded. To sweeten the beverage, used FOS (21%), FOS+Reb B (150 ppm), andFOS+Reb A/Reb B (68 ppm). Descriptive Sensory evaluation was conductedwith 8-13 panelists. Sensory results showed that, with the addition ofReb B, there was enhancement in the sweetness profile of the alcoholicbeverage product. The sweetness onset was earlier in the profile, andthere was an increase in total sweetness and the sweetness profile wasmore rounded. There was a decrease in flavor intensity, bitterlingering, and tartness. FIG. 7b shows the impact of different sweetenersystem on the taste attributes of the beverage.

Example 14 Effect of Reb B on No Fat (NF) Yogurt

Samples were prepared according to formulas outlined below in Table 6.Samples included a full sugar reference, an aspartame reference, a Reb Acontrol and several test samples with a Reb A/Reb B blends. The sampleswere evaluated by a trained panel and screened for overall sweetness,bitterness, astringency, mouth coating, flavor intensity,acidity/tartness, sweet lingering and bitter lingering. Reb A hadbitterness at the end that was not present in samples with Reb A/Reb Bblends. Both Reb A and blends enhanced the acidity of the yogurt. Asignificant improvement of taste was found with the blend of Reb B/RebA, where Reb B ratio to total glycosides (TSG) ranged between 0.5 to50%, more specifically 10% to 40% of Reb B to total glycosides.

TABLE 6 Control 1 2 3 4 5 NF Yogurt (%) 92 99.96 99.94 99.93 99.94 99.94Sugar (%) 8 Aspartame (%) 0.0400 Reb A (%) 0.0250 0.0250 0.0250 NSF02(natural 0.0350 0.0350 0.0350 0.0350 flavor) (%) Reb B (%) 0.0056 0.01400.0090 Reb B/TSG 0 0 0 16% 36% 26% (%)

Example 15 Chocolate Milk

A series of samples were prepared to having sugar-equivalent sweetnessaround 5.5%. The control sample was sweetened with 5.5% sugar the testsamples were sweetened with different ratios of Reb A/Reb as shown inTable 7 below. The samples were evaluated by a trained panel andscreened for overall sweetness, bitterness, astringency, mouth coating,flavor intensity, acidity/tartness, sweet lingering and bitterlingering. Pure Reb A had separate peaks for sweetness, dairy notes, andcocoa notes. The test samples had a very balanced and more roundedsweetness profile with an increasing amount of Reb B content in thesweetener as outlined in the following table. A significant improvementof taste was found with the blend of Reb A/Reb B. where Reb B to totalglycosides ratio ranged between 0.5 to 50, more specifically 10 to 400of Reb B to total glycosides.

TABLE 7 Sample # Ingredient, % Control 1 2 3 4 5 6 Low-fat milk 94.0099.48 99.47 98.47 99.47 99.46 99.45 or Skim milk Sugar 5.50 Cocoa Powder0.50 0.50 0.50 1.50 0.50 0.50 0.50 Reb A 0.0242 0.0242 0.0242 0.01690.0121 Reb B 0.0046 0.0060 0.0157 0.0262 0.0524 Carrageenan 0.01600.0160 0.0160 0.0160 0.0160 0.0160 Total 100 100.0 100.0 100.0 100.0100.0 100.0 Reb B/TSG ratio 0% 16% 20% 48% 68% 100%

Example 16 Table Top Sweeteners

Several table top formulations were developed using different bulkingagents, as examples sugar, fructose, dextrose, maltodextrin, polyols,fibers (inulin, Fibersol-2) etc. Table 8 lists a few examples of theformulation with different ratio of Reb B to total glycosides anddextrose as the bulking agent. Each sample of 1 g wt. was developed forproviding sweetness equivalent to 8-9 grams of sugar. Each sample wastested in coffee by 11 panel members and screened for sweetness, coffeeflavors, bitterness, overall mouthfeel, and residual (lingering)sweetness and bitterness. As the ratio of Reb B to total steviolglycosides increased the overall taste and preference increased. Asignificant improvement of overall sweetness and mouthfeel was foundwith the blend of Reb A/Reb B, where Reb B to total glycosides rangedbetween 1 to 40%, more specifically 7% to 30% of Reb B to totalglycosides.

TABLE 8 Control Sample 1 Sample 2 Sample 3 Reb A (g) 0.0400 0.03360.0300 Reb B (g) 0.0064 0.0100 0.0500 Dextrose (g) 0.9600 0.9560 0.96000.9500 Reb B/TSG (%) 0 16 25 100

Example 17 Baked Goods

The texture and taste profile of baked goods improved with the blend ofReb B and Reb A. Several samples were developed with different ratios ofReb B and Reb A or SG95 (a steviol glycoside product available fromPureCircle Inc.); some examples are shown in the Table 9. The sampleswere evaluated by a trained internal panel and screened for overallsweetness, texture, mouthfeel, mouth coating, flavor intensity, sweetlingering and bitter lingering. The test sample had a better balance ofsweetness and overall acceptability. A significant improvement ofoverall sweetness and mouthfeel was found with the blend of Reb B andReb A or SG95, where Reb B to total glycosides ranged from 0.5% to 40%,more specifically 7% to 30% of Reb B to total glycosides.

TABLE 9 Ingredients (%) Control Sample 1 Sample 2 Sample 3 All-purposeflour 24.42 17.629 17.629 17.629 Sugar 23.14 12.372 12.372 12.372 Wholewheat flour 5.8763 5.8763 5.8763 Maltodextrin 2.1368 2.1368 2.1368Fibersol-2 1.0684 1.0684 1.0684 Modified starch 1.0684 1.0684 1.0684Poppy seeds 1 1.0684 1.0684 1.0684 Baking powder 0.48 1.0684 1.06841.0684 Lemon flavor 0.83 0.886 0.886 0.886 Salt 0.71 0.7479 0.74790.7479 Baking soda 0.3205 0.3205 0.3205 NSF-02 (Natural 0.0635 0.06350.0635 flavor) Reb B 0.025 0.02 SG95 0.0302 0.0302 Reb A 0.005 0.01 WetIngredients Milk, (2% fat) 23.99 27.244 27.244 27.244 Soybean oil 14.7811.753 11.753 11.753 Eggs 9.54 8.5473 8.5473 8.5473 Water 5.342 5.3425.342 Yogurt, Plain 1.6026 1.6026 1.6026 non-fat Lemon juice 0.59 0.6410.641 0.641 Vanilla extract 0.52 0.5342 0.5342 0.5342 Total 100 100 100100 Reb A/TSG (%) 0 0 16 33

Example 18

Black Tea with Peach Flavor

The control tea sample had 7.7% sugar content. The test formulas (Table10) were modified to reduce 70%/a sugar with Reb A or Reb A/B blend(Example 3). The samples were evaluated by a trained panel and screenedfor overall sweetness, sweet onset, mouthfeel, flavor intensity,bitterness, sweet lingering and bitter lingering. Among the testsamples, the sample with Reb A/Reb B had a better balance of sweetness,overall acceptability, earlier onset of sweetness and sugar-like tasteprofile.

TABLE 10 Ingredient (%) Control Reb A/B Reb A Water 91.849 97.271597.27498 Sugar 7.7000 2.2500 2.2500 Black Tea 0.2750 0.2750 0.2750Sodium Citrate 0.0550 0.0550 0.0550 Citric Acid 0.0440 Ascorbic Acid0.0440 0.0440 0.0440 Peach Flavor 0.0330 0.0330 0.0330 Xanthan Gum0.0035 0.0035 Malic Acid 0.0440 0.0440 Reb A/B 0.0240 Reb A 0.0205 TOTAL100 100 100

Example 19 No-Sugar Added 50% Orange Juice Drink

The juice drinks were made with sugar, Reb A/Reb B (Example 3) blend,and Reb A (steviol glycosides) as shown in the Table 11 below. Allsamples had a pH around 3.75. The samples were evaluated by a trainedpanel and screened for overall sweetness, sweet onset, mouthfeel, flavorintensity, bitterness, sweet lingering and bitter lingering. The samplewith Reb A/Reb B was less bitter, more tart, more fresh orange flavor,less astringent and less bitter lingering than that with Reb A. RebA/Reb B was perceived as significantly closer to control in overallflavor profile than Reb A formula.

TABLE 11 Ingredient (%) Control-Sugar Reb A/B Reb A Water 43.940049.9075 49.9160 Orange Juice 50.0000 50.0000 50.0000 Sugar 6.0000Steviol Glycosides 0.0225 0.0215 Flavor 0.0400 0.0700 0.0825 Citric Acid0.0200 TOTAL 100 100 100

Example 20 Lemon-Lime Carbonated Soft Drink

The control carbonated soft drink sample was made with high fructosecorn syrup (HFCS) 42 and other test samples were made with a combinationof HFCS and Reb A/B (Example 3) blend or Reb A to attain 30% lesscalorie (Table 12). The samples were evaluated by a trained panel andscreened for overall sweetness, sweet onset, mouthfeel, flavorintensity, bitterness, sweet lingering and bitter lingering. Productwith Reb A/B feels smoother in mouth and is identified as closer tocontrol than Reb A formula. It has less of a sharp sweetness peak thanReb A and reduced terpene notes compared to Reb A. The overall flavor isless altered compared to Reb A.

TABLE 12 Ingredient (%) Control Reb A/B Reb A Water 84.85 89.30 89.30HFCS 42 14.83 10.38 10.38 Reb A 0.0049 Reb A/B 0.0058 Xanthan gum 0.00500.0050 Sodium Benzoate 0.0263 0.0263 0.0263 Potassium Citrate 0.02630.0263 0.0263 Citric Acid 0.1500 0.1433 0.1433 Malic Acid 0.0050 0.0050Flavor 0.1110 0.1110 0.1110

Example 21 Peppermint Mouthwash

A commercial unsweetened (no sugar or sweetener) peppermint mouthwashproduct (Brand: The Natural Dentist) was used to determine the effect ofReb B with other sweetener, as example maltitol, a sugar alcohol on thesensory profile of mouthwash. Mouthwash samples were sweetened withMaltitol (6.44%), Maltitol+Reb B (50 ppm), Maltitol+Reb B (150 ppm).Descriptive sensory evaluation was conducted with 8 panelists. Theyevaluated several attributes including sweet onset, total sweetness,bitterness, rounded sweetness, flavor intensity, astringency, mouthcoating, sweet lingering, bitter lingering, aftertaste mouth coating,tartness. Sensory results showed that the addition of Reb B created anearlier sweetness onset, and higher total sweetness. All otherattributes were generally unchanged.

Although the invention and its advantages have been described in detail,it should be understood that various changes, substitutions andalterations can be made herein without departing from the spirit andscope of the invention as defined by the appended claims. Moreover, thescope of the application is not intended to be limited to the particularembodiments of the invention described in the specification. As oneskilled in the art will readily appreciate from the disclosure of theinvention, the compositions, processes, methods and steps, presentlyexisting or later to be developed that perform substantially the samefunction or achieve substantially the same result as the correspondingembodiments described herein are encompassed by the scope of theinvention.

1. A sweet ingestible composition comprising Rebaudioside B and at leastone sweetener, selected from the group consisting of steviol glycosides,stevia extracts, natural sweeteners, glycosylated terpenoid sweeteners,synthetic high intensity sweeteners, oligosaccharides, caloricsweeteners, and combinations thereof, wherein said at least onesweetener is present in the composition at a concentration above 2%sucrose equivalent sweetness, and Rebaudioside B have a purity of atleast 95% wt/wt on anhydrous basis is present at a concentration from 10to 300 ppm.
 2. The sweet ingestible composition of claim 1, comprisingone or more steviol glycosides selected from the group consisting ofstevioside, Rebaudioside A, Rebaudioside C, Rebaudioside D, RebaudiosideE, Rebaudioside F, dulcoside A, steviolbioside, rubusoside, andcombinations thereof.
 3. The sweet ingestible composition of claim 1,comprising one or more natural sweeteners selected from the groupconsisting of Luo Han Guo extract, Mogrosides, Mogroside V,Glycyrrhizin, Thaumatin, Brazzein, and combinations thereof.
 4. Thesweet ingestible composition of claim 1, comprising one or moreglycosylated terpenoid sweeteners selected from the group consisting ofa glucosylated steviol glycoside, a glucosylated mogroside andcombinations thereof.
 5. The sweet ingestible composition of claim 1,comprising at least one synthetic high intensity sweetener selected fromthe group consisting of aspartame, saccharin, sucralose, Acesulfame-K,neotame and combinations thereof.
 6. The sweet ingestible composition ofclaim 1, comprising at least one sugar alcohol selected from the groupconsisting of erythritol, maltitol, sorbitol, xylitol and combinationsthereof.
 7. The sweet ingestible composition of claim 1, comprising atleast one oligosaccharide selected from the group consisting offructo-oligosaccharides, inulin, inulo-oligosaccharides, polydextrose,and malto-oligosaccharides and combinations thereof.
 8. The sweetingestible composition of claim 1, comprising at least one caloricsweetener selected from the group consisting of sugar, invert-sugar,fructose, dextrose, maltose, lactose, corn syrup, HFCS, and combinationsthereof.
 9. A food or beverage product having sweetness, enhancedsweetness, enhanced flavor, or a combination thereof, comprising thesweet ingestible composition of claim
 1. 10. A drug, pharmaceutical orcosmetic preparation comprising the sweet ingestible composition ofclaim
 1. 11. A sweetener comprising the sweet ingestible composition ofclaim
 1. 12. The sweet ingestible composition of claim 1, furthercomprising a flavoring agent.
 13. The sweet ingestible composition ofclaim 1, further comprising a food ingredient selected from the groupconsisting of acidulants, organic and amino acids, coloring agents,bulking agents, modified starches, gums, texturizers, preservatives,antioxidants, emulsifiers, stabilizers, thickeners, gelling agents, andcombinations thereof
 14. (canceled)